This application is a 371 of PCT/FR99/01716, filed Jul. 13, 1999.
The present invention relates to pharmaceutical compositions comprising 2-quinolones or derivative compounds.
A cancer is a disorder of the somatic genes in the course of which genetic dysfunctions are amplified as the tumoral process progresses from the state of precancerous lesion to that of malignant transformation, the cancer tumor becoming metastatic and often resistant to cytotoxic medicines.
Despite the very considerable efforts made in all the developed countries, in particular through experimental and clinical research programs, mortality due to various cancers (solid tumors and hematological neoplasties) remains unacceptably high. In many countries, cancer is the second most common cause of death, just after cardiovascular diseases.
In terms of newly diagnosed cancers, the distribution between solid tumors and hematological neoplasties (bone marrow, blood, lymphatic system) shows that 9 out of 10 cancers are solid tumors. In contrast with what is observed in hematological oncology (therapeutic successes in 40% to 90% of cancers of blood cells), only a small number of advanced or disseminated solid tumors respond to chemotherapy treatments alone. It is partly for this reason that the overall death by cancer grew in the USA between 1973 and 1992.
Unfortunately, it is not certain that this tendency may be reversed merely by the appearance, alongside the established chemotherapy arsenal, of new antitumor medicines such as taxanes (paclitaxel and docetaxel) which interfere with the formation of microtubules (W. P. McGuire et al., Am. Intern. Med., 1989), topoisomerase I inhibitors derived from camptothecin (topotecan and irinotecan), vinorelbine (novel alkaloid derived from periwinkle), gemcitabine (novel cytotoxic antimetabolite), raltitrexed (thymidylate synthetase inhibitor) and miltefosine (first representative of the alkyl-lysophospholipid family). These treatments are added, either as a first line treatment or as a second line treatment, to medicines whose specific activity is now well recognized, such as doxorubicin, cysplatin, vincristine, methotrexate and 5-fluorouracil.
One of the most difficult current problems in anticancer chemotherapy is due to the fact that many populations of malignant cells show considerable resistance to the established cytotoxic substances. Usually, this situation results from the existence of multi-drug-resistance genes or from the frequency of genetic mutations in certain types of tumors. Thus, the treatment of cancers requires novel approaches, complementary to those currently used, and designed to better combat the extension and heterogeneity of the tumor load and the acquisition of xe2x80x9ccytotoxic multidrugxe2x80x9d resistance.
Among these novel approaches, some are already promising. This is the case for the induction of apoptosis, the inhibition of tumor angiogenesis and of metastatic processes, not to mention gene therapy or immunotherapy.
The inventors have become interested in a different approach. The desired objective was to make the population of tumor cells more sensitive to the standard anticancer treatments in order to achieve a twofold benefit:
1) to increase the cytotoxic activity and thus the efficacy, and
2) to reduce the frequency and severity of certain side effects by means of reducing the dosage which might follow the induction of the increase in the antitumor efficacy.
It is this strategy which underlies the discovery of compositions capable of inducing a highly significant increase in the cytotoxic activity of tested anticancer medicines. These compositions have the capacity either of stimulating the recruitment of clonogenic cells in the tumor, thus making it more sensitive to the conventional treatment with cytotoxic agents, or of inhibiting the proliferation of clonogenic cells, thus contributing toward the regression of the tumor.
A subject of the present invention is thus the use, in the treatment of cancers with at least one antitumor agent chosen from cytotoxic agents, of a compound having activity on the proliferation of clonogenic cells in tumors, chosen from the compounds of formulae: 
in which:
X is chosen from xe2x95x90O, xe2x95x90S and xe2x95x90Nxe2x80x94NHxe2x80x94R7, R7 being a phenyl or pyridyl group,
R1, R2, R3 and R4 are chosen, independently of each other, from H, OH, a C1-C4 alkyl group, a C1-C4 alkoxy group, a group xe2x80x94OCOxe2x80x94R8, R8 being a C1-C4 alkyl group, and a group derived from a saccharide, at least one of the substituents R1, R2, R3 or R4 being other than H, and R2 and R3 together possibly forming a methylenedioxy group,
R5 is a phenyl group or a phenyl group substituted 1 to 3 times with groups chosen from H, OH, a C1-C4 alkoxy group, a group xe2x80x94OCOR8, a phenyl (C1-C4 alkoxy) group, a group xe2x80x94Oxe2x80x94SO2xe2x80x94Rxe2x80x28, Rxe2x80x28 being a C1-C4 alkyl group or a CF3 group, and a group derived from a saccharide,
R6 is chosen from H, a C1-C4 alkyl group, a group xe2x80x94COxe2x80x94R9 and a group xe2x80x94Axe2x80x94R10,
R6a is chosen from a C1-C4 alkyl group, a group xe2x80x94COxe2x80x94R9 and a group xe2x80x94Axe2x80x94R10,
R9 being a C1-C4 alkyl group,
A being a C1-C4 alkylene group,
R10 being chosen from 5- or 6-membered heterocyclic groups containing 1 to 4 hetero atoms chosen from oxygen, sulfur and nitrogen, the CN group, a group xe2x80x94COOR11, xe2x80x94CONR12R13, a group xe2x80x94NR14R15 and a group xe2x80x94COR16, R11, R12, R13, R14, R15 and R16 being chosen independently from a hydrogen atom, a C1-C4 alkyl group and a phenyl(C1-C4 alkyl) group,
R4 and R6 together also possibly forming a xe2x80x94COxe2x80x94CH2xe2x80x94CH2xe2x80x94 group.
The cytotoxic agents may be used at their usual dose and, in this case, their efficacy is improved, or at lower doses given the increase in their antitumor efficacy.
In one preferred embodiment, the compound used is a compound of formula (I) in which:
R1 is a C1-C4 alkoxy group
R2 is a hydrogen atom
R3 is a C1-C4 alkoxy group
R4 is a hydrogen atom,
and in particular a compound of formula (I) in which:
R5 is a 4-(C1-C4 alkoxy)phenyl group,
and most particularly a compound of formula (I) in which:
R1 is a methoxy group,
R3 is a methoxy group, and
R5 is a 4-methoxyphenyl group.
It has also been discovered that at least some of the compounds of formula (I) had antitumor activity themselves.
A subject of the present invention is also a composition having activity on the proliferation of clonogenic cells in tumors by interfering with the generation of clonogenic cells, either by stimulating proliferation and recruitment, or by inhibiting proliferation, and which comprises an effective amount of a compound of formulae: 
in which:
X is chosen from xe2x95x90O, xe2x95x90S and xe2x95x90Nxe2x80x94NHxe2x80x94R7, R7 being a phenyl or pyridyl group, R1, R2, R3 and R4 are chosen, independently of each other, from H, OH, a C1-C4 alkyl group, a C1-C4 alkoxy group, a group xe2x80x94OCOxe2x80x94R8, R8 being a C1-C4 alkyl group, and a group derived from a saccharide, at least one of the substituents R1, R2, R3 or R4 being other than H, and R2 and R3 together possibly forming a methylenedioxy group,
R5 is a phenyl group or a phenyl group substituted 1 to 3 times with groups chosen from H, OH, a C1-C4 alkoxy group, a group xe2x80x94OCOR8, a phenyl (C1-C4 alkoxy) group, a group xe2x80x94Oxe2x80x94SO2xe2x80x94Rxe2x80x28, Rxe2x80x28 being a C1-C4 alkyl group or a CF3 group, and a group derived from a saccharide,
R6 is chosen from H, a C1-C4 alkyl group, a group xe2x80x94COxe2x80x94R9 and a group xe2x80x94Axe2x80x94R10,
R6a is chosen from a group xe2x80x94COxe2x80x94R9 and a group xe2x80x94Axe2x80x94R10,
R9 being a C1-C4 alkyl group,
A being a C1-C4 alkylene group,
R10 being chosen from 5- or 6-membered heterocyclic groups containing 1 to 4 hetero atoms chosen from oxygen, sulfur and nitrogen, the CN group, a group xe2x80x94COOR11, xe2x80x94CONR12R13, a group xe2x80x94NR14R15 and a group xe2x80x94COR16,
R11, R12, R13, R14, R15 and R16 being chosen independently from a hydrogen atom, a C1-C4 alkyl group and a phenyl(C1-C4 alkyl) group,
R4 and R6 together also possibly forming a xe2x80x94COxe2x80x94CH2xe2x80x94CH2xe2x80x94 group.
A subject of the present invention is also novel compounds, namely compounds of formulae: 
in which:
X is chosen from xe2x95x90O, xe2x95x90S and xe2x95x90Nxe2x80x94NHxe2x80x94R7, R7 being a phenyl or pyridyl group,
R1, R2, R3 and R4 are chosen, independently of each other, from H, OH, a C1-C4 alkyl group, a C1-C4 alkoxy group, a group xe2x80x94OCOxe2x80x94R8, R8 being a C1-C4 alkyl group, and a group derived from a saccharide, at least one of the substituents R1, R2, R3 or R4 being other than H, and R2 and R3 together possibly forming a methylenedioxy group,
R5 is a phenyl group or a phenyl group substituted 1 to 3 times with groups chosen from H, OH, a C1-C4 alkoxy group, a group xe2x80x94OCOR8, a phenyl (C1-C4 alkoxy) group, a group xe2x80x94Oxe2x80x94SO2xe2x80x94Rxe2x80x28, Rxe2x80x28 being a C1-C4 alkyl group or a CF3 group, and a group derived from a saccharide,
R6 is chosen from H, a C1-C4 alkyl group, a group xe2x80x94COxe2x80x94R9 and a group xe2x80x94Axe2x80x94R10,
R6a is chosen from a group xe2x80x94COxe2x80x94R9 and a group xe2x80x94Axe2x80x94R10,
R9 being a C1-C4 alkyl group,
A being a C1-C4 alkylene group,
R10 being chosen from 5- or 6-membered heterocyclic groups containing 1 to 4 hetero atoms chosen from oxygen, sulfur and nitrogen, the CN group, a group xe2x80x94COOR11, xe2x80x94CONR12R13, a group xe2x80x94NR14R15 and a group xe2x80x94COR16,
R11, R12, R13, R14, R15 and R16 being chosen independently from a hydrogen atom, a C1-C4 alkyl group and a phenyl(C1-C4 alkyl) group,
R4 and R6 together also possibly forming a xe2x80x94COxe2x80x94CH2xe2x80x94CH2xe2x80x94 group,
with the exclusion of the compounds in which X=O, R6=H and two of the substituents R1, R2, R3 and R4 are OH or OCH3.
In the chemotherapy treatment of cancers with cytotoxic agents, the compounds of formulae (I) and (Ia) can be administered at the start of the chemotherapy treatments either once or over several days at the start of these treatments (for example for 5 to 7 days) and, depending on the chemotherapy protocol, at the start of each treatment cycle (for example for 2 to 5 days) in the course of each cure.
The compounds of formulae (I) and (Ia) are advantageously administered by infusion (generally in 1 to 3 hours) at doses of from 5 to 50 mg/kg/day or 200 to 2000 mg/m2/day.
In order to obtain a maximal effect on the production (inhibition or stimulation) of clonogenic cells, the compounds of formulae (I) and (Ia) should be administered such that the tissue concentrations obtained are as high as can possibly be envisaged.
For the treatment protocols in the acute phases of the cures, the intravenous route is preferred, using:
ready-to-use infusion solutions (bags, bottles, etc.) intended to be administered without any modification, by intravenous infusion using an infusion line and at the recommended rate:
lyophilizates to be redissolved for intravenous infusion using pharmaceutical solutions known to those skilled in the art;
for maintenance treatments, it is also possible to envisage the oral route when the chemotherapy treatment favors the oral administration of cytostatic agents. To this end, lozenges (for oral or perlingual absorption), immediate-release or delayed-release tablets, oral solutions, suspensions, granules, gel capsules, etc. may be used.
The cytotoxic agents may be chosen from:
i) intercalating agents, in particular doxorubicin (Adriamycin), daunorubicin, epirubicin, idarubicin, zorubicin, aclarubicin, pirarubicin, acridine, mitoxanthrone, actinomycin D, eptilinium acetate;
ii) alkylating agents chosen from platinum derivatives (cisplatin, carboplatin, oxaliplatin);
iii) a compound chosen from the other groups of alkylating agents:
cyclophosphamide, ifosfamide, chlormetrine, melphalan, chlorambucil, estramustine,
busulfan, mitomycin C,
nitrosoureas: BCNU (carmustine), CCNU (lomustine), fotemustine, streptozotocin,
triazines or derivatives: procarbazine, dacarbazine,
pipobroman,
ethyleneimines: altretamine, triethylene-thio-phosphoramide,
iv) a compound chosen from the other groups of anti-metabolic agents:
antifolic agents: methotrexate, raltitrexed,
antipyrimidine agents: 5-fluorouracil (5-FU), cytarabine (Ara-C),
hydroxyurea
antipurine agents: purinethol, thioguanine, pentostatin, cladribine,
cytotoxic nucleoside synthesis inducers: gemcitabine,
v) a compound chosen from the other groups of tubulin-affinity agents,
vinca alkaloids which disrupt the mitotic spindle: vincristine, vinblastine, vindesine, navelbine,
agents which block the depolymerization of the mitotic spindle: paclitaxel, docetaxel,
agents which induce DNA cleavage by inhibition of topoisomerase II: etoposide, teniposide,
topoisomerase I inhibitors which induce DNA cleavage: topotecan, irinotecan,
vi) a DNA splitting or fragmenting agent, such as bleomycin,
vii) one of the following compounds: plicamycin, L-asparaginase, mitoguazone, dacarbazine,
viii) an anticancer progestative steroid; medroxy-progesterone, megestrol,
ix) an anticancer estrogen steroid: diethylstilbestrol; tetrasodium fosfestrol,
x) an antiestrogen agent: tamoxifen, droloxifen, raloxifen, aminoglutethimide,
xi) a steroidal antiandrogenic agent (eg cyproterone) or a non-steroidal antiandrogenic agent (flutamide, nilutamide).
In particular, the compounds of formulae (I) and (Ia) may be combined with all the major treatments with cytotoxic agents used in solid tumor polychemotherapies, such as:
doxorubicin
alkylating agents: oxazophorines (cyclophosphamide, ifosfamide, chlorambucil, melphalan)
nitrosoureas
mitomycin C
antimetabolites such as methotrexate, 5-FU, Ara-C, capecitabine
agents which interfere with tubulin: vinca alkaloids (vincristine, vinblastine, vindesine, navelbine), taxoids (paclitaxel, docetaxel), epipodophyllotoxin derivatives (etoposide, teniposide)
bleomycin
topoisomerase I inhibitors: topotecan, irinotecan.
Similarly, the compounds of formulae (I) and (Ia) may be combined with the treatments with the major cytotoxic agents used in oncohematology for the treatment of blood cancers:
Hodgkin""s disease: cyclophosphamide, mechlorethamine, chlorambucil, melphalan, ifosfamide, etoposide, doxorubicin, daunorubicin;
acute leukemias: methotrexate, 6-mercaptopurine, cytarabine, vinblastine, vincristine, doxorubicin, daunorubicin, L-asparaginase;
non-Hodgkin""s malignant lymphomas: mechlorethamine, chlorambucil, cyclophosphamide, melphalan, ifosfamide, methotrexate, cytarabine, vinblastine, vincristine, etoposide, doxorubicin, daunorubicin, carmustine, lomustine, cisplatin;
chronic lymphoid leukemias: mechlorethamine, chlorambucil, cyclophosphamide, melphalan, ifosfamide.
In general, the compounds of formula (I) may be prepared according to the following reaction schemes: 
A reagent of the type XR6 in which X=I, Br or Cl may be used as alkylating agent.
As a variant, a compound CH2=CHxe2x80x94R can be used to attach a group R6=xe2x80x94CH2xe2x80x94CH2xe2x80x94R (corresponding to the group xe2x80x94Axe2x80x94R10 defined above). 
In addition, it is possible to convert some or all of the alkoxy groups into hydroxyl groups according to known methods. Similarly, the hydroxyl groups may be converted into ester or sulfonate according to the known methods.
Similarly, it is possible to convert a group xe2x80x94Axe2x80x94COOR11, in which R11 is an alkyl or phenylalkyl group, into a group xe2x80x94Axe2x80x94COOH and to convert a group xe2x80x94Axe2x80x94COOH into a group xe2x80x94Axe2x80x94CONR12R13, according to known methods.
The compounds in which R4 and R6 form a xe2x80x94COxe2x80x94CH2xe2x80x94CH2xe2x80x94 group can be obtained by cyclization of a compound in which R4=H and R6=xe2x80x94CH2xe2x80x94CH2xe2x80x94COOH.